Utility of flow cytometric PAX5 protein detection for the diagnosis of B-cell acute lymphoblastic leukemia
Issued Date
2026-01-01
Resource Type
ISSN
14346621
eISSN
14374331
Scopus ID
2-s2.0-105028619659
Journal Title
Clinical Chemistry and Laboratory Medicine
Rights Holder(s)
SCOPUS
Bibliographic Citation
Clinical Chemistry and Laboratory Medicine (2026)
Suggested Citation
Pootrakul T., Khiankaew B., Boonsakan P., Paisooksantivatana K. Utility of flow cytometric PAX5 protein detection for the diagnosis of B-cell acute lymphoblastic leukemia. Clinical Chemistry and Laboratory Medicine (2026). doi:10.1515/cclm-2025-1647 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/114836
Title
Utility of flow cytometric PAX5 protein detection for the diagnosis of B-cell acute lymphoblastic leukemia
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Abstract
Accurate B-lineage assignment in acute leukemia is critical for therapeutic decisions. While nuclear expression of PAX5 protein (nPAX5) is a highly reliable marker for B-cell differentiation, its conventional assessment by time-consuming immunohistochemistry (IHC) is not ideal for rapid diagnosis. This study aimed to validate the utility of flow cytometric (FCM) detection of nPAX5 for the diagnosis of B-lymphoblastic leukemia (B-ALL), potentially enhancing diagnostic efficiency. A retrospective study was conducted, comparing nPAX5 expression by FCM and IHC in 125 bone marrow biopsies (57 B-ALL, 12 T-ALL, 56 AML). Further diagnostic performance analysis, using ROC analysis of the geometric mean fluorescence intensity ratio (Blast/Normal T-cell), was performed for nPAX5, cCD22, and cCD79a across a larger cohort of 538 acute leukemia cases (123 B-ALL, 29 T-ALL, 386 AML). FCM and IHC for PAX5 expression showed 100% concordance across the 125 cases (57/57 B-ALL positive; 0/12 T-ALL negative). The single PAX5-positive AML case harbored the t(8;21) translocation. ROC analysis demonstrated excellent diagnostic performance for both cCD79a (AUC 0.980) and nPAX5 (AUC 0.955). At optimal criteria, cCD79a achieved 100.00% specificity and 91.06% sensitivity. nPAX5 showed strong utility, matching the 91.06% sensitivity with 89.45% specificity (criterion >6.30). cCD22 exhibited moderate discriminatory power (AUC 0.807). Flow cytometric nuclear PAX5 (nPAX5) demonstrated 100% agreement with IHC and exhibited excellent diagnostic accuracy (AUC 0.955, 91.06% sensitivity). These compelling results validate nPAX5 as a reliable and powerful FCM marker, supporting its immediate adoption for rapid and efficient B-lineage assignment in acute leukemia diagnostics.